Potential role for gut microbiota in cell wall digestion and glucoside detoxification in Tenebrio molitor larvae

Tenebrio molitor larvae were successfully reared free of cultivatable gut lumen bacteria, yeasts and fungi using two approaches; aseptic rearing from surface sterilized eggs and by feeding larvae with antibiotic-containing food. Insects were reared on a rich-nutrient complete diet or a nutrient-poor refractory diet. A comparison of digestive enzyme activities in germ free and conventional insects containing a gut microbiota did not reveal gross differences in enzymes that degrade cell walls from bacteria (lysozyme), fungi (chitinase and laminarinase) and plants (cellulase and licheninase). This suggested that microbial-derived enzymes are not an essential component of the digestive process in this insect. However, more detailed analysis of T. molitor midgut proteins using an electrophoretic separation approach showed that some digestive enzymes were absent and others were newly expressed in microbiota-free larvae. Larvae reared in antibiotic-containing refractory wheat bran diet performed poorly in comparison with controls. The addition of saligenin, the aglycone of the plant glucoside salicin, has more deleterious effects on microbiota-free larvae than on the conventionally reared larvae, suggesting a detoxifying role of midgut microbiota. Analysis of the volatile organic compounds released from the faecal pellets of the larvae shows key differences in the profiles from conventionally reared and aseptically reared larvae. Pentadecene is a semiochemical commonly found in other beetle species. Here we demonstrate the absence of pentadecene from aseptically reared larvae in contrast to its presence in conventionally reared larvae. The results are discussed in the light of the hypothesis that microbial products play subtle roles in the life of the insect, they are involved in the digestion of refractory food, detoxification of secondary plant compounds and modify the volatile profiles of the insect host.     

Studies on the diapause of Trogoderma granarium: Effects of juvenile hormone analogues on growth and metamorphosis

Trogoderma granarium larvae reared at 30°C on wheat flour containing appropriate concentrations of farnesyl methyl ether (FME) resembled normal diapause larvae in their growth characteristics and reactions to diapauseterminating conditions. When these larvae were transferred to 37°C either without food or with normal food, normal adults resulted. But when the diapause-like state was broken at 37°C in the presence of the original FME-containing food the insects either died before pupal-adult ecdysis or gave abnormal adults. At 35°C FME did not inhibit the larval-pupal ecdysis, but only a few adults emerged and these were abnormal.Farnesol did not inhibit metamorphosis at 30°C and caused less abnormality than did FME.Based on these results, the hypothesis that the density-dependent diapause of this insect is caused by the presence, in faecal pellets, and thereby in contaminated food, of juvenile hormone (JH) analogues is rejected, but the possibility that diapause results from a high physiological level of JH brought about by causes other than the presence of JH analogues in the environment is examined.     

Preimaginal conditioning does not affect oviposition preference in the diamondback moth

Abstract.  1. In holometabolous insects, learning has been demonstrated in both larval and adult stages. Whether learning can be retained through metamorphosis from larva via pupa to adult has long been a subject of debate. The present study is designed to distinguish between preimaginal and imaginal conditioning in the diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae) using oviposition preference tests on females exposed to various types of learning experiences during immature and adult stages.
2. Cohorts of test insects were reared from egg to pupa on an artificial diet, or on one of two host plants, Chinese cabbage, Brassica campestris L. ssp. pekinensis , and common cabbage, Brassica oleracea L. var. capitata . The ensuing females reared on the three kinds of food showed similar oviposition preference between the two plants. A brief experience of the less preferred host, common cabbage, by adults slightly increased their preference for this plant.
3. Cohorts of test insects were reared from egg to pupa on an artificial diet with or without the addition of a neem-based oviposition deterrent (Neemix^® 4.5). Larval feeding experience did not alter oviposition response to the deterrent. However, emergence conditioning and early adult learning, achieved through experience of a residue of the deterrent carried over from the larval food on pupal cuticle and cocoons, altered oviposition preference significantly.
4. The combined results revealed no evidence of preimaginal conditioning in this insect but a strong effect of emergence conditioning and early adult learning on oviposition preference.     

Effect of different artificial diets on diapause induction under controlled temperature and photoperiod in the silkworm, Bombyx mori L.

ABSTRACT. Embryonic diapause of the silkworm, Bombyx mori , is generally induced by temperature and photoperiod during the egg stage of the previous generation and not in the larval stage. However, when silkworm larvae are reared on an artificial diet instead of mulberry leaves, their diapause is strongly affected by temperature and photoperiod experienced in the larval stage, with a distinct long-day response for diapause induction. Moreover when larvae which have been reared on artificial diet under long-day condition are fed mulberry leaves even for a short period of time, most of the resultant female adults lay diapause eggs. These results suggest that the photoperiodic response of larvae for diapause induction may be strongly suppressed by some components in mulberry leaves.     

Temperature and food quality effects on growth,consumption and post-ingestive utilization efficiencies of the forest tent caterpillar Malacosoma disstria (Lepidoptera: Lasiocampidae)

Temperature and food quality can both influence growth rates, consumption rates, utilization efficiencies and developmental time of herbivorous insects. Gravimetric analyses were conducted during two consecutive years to assess the effects of temperature and food quality on fourth instar larvae of the forest tent caterpillar Malacosoma disstria Hübner. Larvae were reared in the laboratory at three different temperatures (18, 24 and 30 degrees C) and on two types of diet; leaves of sugar maple trees Acer saccharum Marsh. located at the forest edge (sun-exposed leaves) or within the forest interior (shade-exposed leaves). In general, larvae reared at 18 degrees C had lower growth rates and lower consumption rates than larvae reared at the warmer temperatures (24 and 30 degrees C). Moreover, the duration of the instar decreased significantly with increasing temperatures. Type of diet also affected the growth rates and amount of food ingested by larvae but did not affect the duration of the instar. Larvae fed sun-exposed leaves consumed more food and gained higher biomasses. Values of approximate digestibility and efficiency of conversion of ingested food were also higher when larvae were fed sun-exposed leaves. Higher growth rates with increasing temperatures were primarily the result of the shorter stadium duration. The higher growth rates of larvae fed sun-exposed leaves were possibly the result of stimulatory feeding and consequently greater food intake and also a more efficient use of food ingested. This study suggests that the performance of M. disstria caterpillars could be enhanced by warmer temperatures and higher leaf quality.     

Role of granivory and insectivory in the life cycle of the carabid beetle Amara similata

1. The cereal field carabid beetle Amara similata was selected to elucidate the role of seeds and insects as food sources. Three experiments were performed to rank different weed seeds and insects in terms of food value: (i) fecundity in relation to adult diet, (ii) larval survival in relation to diet and (iii) larval survival in relation to parental diet.
2. Seeds were found to be of high value and insects of low value both for adults and larvae. Adding insects to a seed diet gave no significant improvement. The value of single species of seeds varied within adults and larvae, but also between adults and larvae. This is the first report that Amara larvae are granivorous.
3. The value of different insects varied by species. Cereal aphid species were of the lowest value both for adult and larval beetles. On a diet of mixed insects, the adults were able to produce a low number of eggs but the larvae all died before pupation.
4. There was no simple relation between survival of the larvae and the quality of the parents' diet.     

重金属污染影响植食性昆虫的研究进展

重金属污染是世界各国面临的最为棘手的问题之一,对生态系统和食品安全构成了严重威胁。作为生态系统中食物链和食物网的重要环节,植食性昆虫是环境中重金属迁移、积累的重要媒介,其因重金属污染而受到的影响引起了大家的关注。本文综述了从2007至2018年重金属污染对植食性昆虫影响的研究进展。昆虫受重金属胁迫的研究途径有人工饲料添加、野外田间暴露、“土壤-植物-昆虫”食物链传递以及体外注射等。积累在植食性昆虫体内的过量重金属可导致其存活率、繁殖力和种群增长率降低,生长发育迟缓。重金属污染对植食性昆虫的生理生化毒性包括细胞超微结构破坏和DNA损伤,体内能量物质含量降低,酶活性、基因表达改变等。植食性昆虫会通过重金属硫蛋白、解毒酶活性的诱导等机制抵御重金属的毒害,从而对低浓度、长期重金属暴露产生生态适应性,甚至提高对其他逆境(如农药等)的耐受性。     

Phylloplane bacteria increase the negative impact of food limitation on insect fitness

1. When populations of herbivorous insects increase in density, they can alter the quantity or quality of their food. The impacts of diet‐related stressors on insect fitness have been investigated singly, but not simultaneously. 2. Foliage quantity and quality of red alder, Alnus rubra, were manipulated together with the presence of non‐entomopathogenic phylloplane bacteria to investigate their impacts, singly and in combination, on survival, pupal mass, growth rate, fecundity and egg quality of a cyclic forest insect, the western tent caterpillar, Malacosoma californicum pluviale. 3. Food limitation (half food) had strong negative impacts on all life‐history traits. When the larvae were fed continuously, however, neither ingesting phylloplane bacteria nor eating leaves from damaged branches (induced foliage) affected survival. In the half‐food treatment, ingesting bacteria further increased mortality, while feeding on induced foliage improved survival. 4. Growth rate and pupal mass of both sexes were reduced for larvae with food limitation compared with continuously fed insects and this was exacerbated when the larvae also ate bacteria‐treated leaves. A combination of bacteria and induced foliage also reduced larval growth rate by 5% in the full‐food treatment. 5. Fecundity (eggs per egg mass) was 2.7 times greater in full‐food than in food‐limited treatments but neither phylloplane bacteria nor plant induction had an effect. Insects fed induced foliage produced smaller eggs. Overall, there was no evidence of a three‐way interaction between the three stressors, although there were negative synergistic effects, primarily between food limitation and the ingestion of phylloplane bacteria.     

Insect responses to plant water deficits. II. Effect of water deficits in soybean plants on the growth and survival of Mexican bean beetle larvae

Abstract. 1. We examined the effect of water deficits in soybean, Glycine max [L.] Merr., on the growth and survival of Mexican bean beetle larvae, Epilachna varivestis Mulsant (Coleoptera: Coccinellidae).
2. Larvae were reared under growth chamber, glasshouse, and field conditions on foliage from plants that were either well-watered or subjected to water deficits.
3. Larval survival, growth rate and pupal weight were reduced, and development time was increased when larvae were reared on foliage from plants subjected to water deficits. These results are contrary to White's (1974) hypothesis that water deficits in plants cause increased growth and survival of herbivorous insects.
4. Changes in foliage chemistry caused by water deficits, possibly in the concentration of free amino acids, are the most likely causes of the observed effects on growth rate and development time. However, under glasshouse and field conditions, physical changes in foliage and/or the foliage's environment that accompany water deficits are also important.     

Geographic and parental influences on diapause by a polyphagous insect herbivore

1 Facultative diapause permits insects to initiate an additional generation when conditions are favourable, or to enter diapause when they are not. Although the principle environmental cues for diapause induction are often temperature and photoperiod, we demonstrated recently that the quality of host plant consumed by the larvae of Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae) influences the proportion of larvae that enter diapause and therefore voltinism. 2 Previous work has also suggested that non‐Mendelian parental effects influence diapause by C. rosaceana larvae, but the mechanism was not established. Here, we report further evidence for parental effects on diapause and examine whether parental diet influences diapause induction. 3 We collected larvae of C. rosaceana from six sites at three latitudes in the Province of Québec. There was an increase in the proportion of larvae entering diapause with increasing latitude. Rearing larvae under identical conditions on artificial diet for two generations significantly reduced variation in diapause among populations within a given latitude, and supported the existence of parental effects that influence offspring diapause. 4 Experiments with different host plants and high‐ vs. low‐quality artificial diets provided no evidence that the parental effects on offspring diapause were based on the quality of food consumed by parents. Diapause in C. rosaceana appears influenced by a hierarchy of cues, including temperature, photoperiod, current larval diet and parental environment.     

Pre‐wintering conditions and post‐winter performance in a solitary bee: does diapause impose an energetic cost on reproductive success?

1. Diapause is a dynamic process of low metabolic activity that allows insects to survive periods of harsh conditions. Notwithstanding the lowered metabolism, and because diapausing insects have no access to food, diapause has an energetic cost that may affect post‐diapause performance. 2. Previous studies on the solitary bee Osmia lignaria have shown that prolonged pre‐wintering periods (the time during which individuals already in diapause remain at warm temperatures) are associated with elevated lipid consumption, fat body depletion, and body weight loss. The present study investigated whether prolonged pre‐wintering also affects reproduction, i.e. whether the costs associated with diapause could have an effect on post‐diapause performance in this species. 3. Females were exposed to a range of pre‐wintering conditions, and ovary development and individual post‐wintering performance were monitored throughout their adult life span. 4. No evidence of an effect of pre‐wintering duration on post‐diapause reproductive success was found. Expected differences in the timing of establishment were not observed because ovary maturation was, surprisingly, not arrested during pre‐wintering. Prolonged pre‐wintering duration did not result in decreased life span, probably because emerging females could rapidly replenish their metabolic reserves through feeding. However, there was a very strong effect of the duration of the pre‐emergence period on the likelihood of nest establishment. 5. Longevity, the main factor determining fecundity in Osmia, is subjected to high levels of intrinsic variability, even among females of similar size exposed to identical conditions during development and nesting.     

Fitness costs associated with Cry1F resistance in the European corn borer

Crops producing insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) are widely planted to manage insect pests. Bt crops can provide an effective tool for pest management; however, the evolution of Bt resistance can diminish this benefit. The European corn borer, Ostrinia nubilalis Hübner, is a significant pest of maize and is widely managed with Bt maize in the Midwest of the United States. When Bt crops are grown in conjunction with non‐Bt refuges, fitness costs of Bt resistance can delay the evolution of resistance. Importantly, fitness costs often vary with ecological factors, including host‐plant genotype and diapause. In this study, we examined fitness costs associated with Cry1F resistance in O. nubilalis when insects were reared on three maize lines. Fitness costs were tested in two experiments. One experiment assessed the fitness costs when Cry1F‐resistant and Cry1F‐susceptible insects were reared on plants as larvae and experienced diapause. The second experiment tested resistant, susceptible and F1 heterozygotes that were reared on plants but did not experience diapause. Despite some evidence of greater adult longevity for Cry1F‐resistant insects, these insects produced fewer fertile eggs than Cry1F‐susceptible insects, and this occurred independent of diapause. Reduced fecundity was not detected among heterozygous individuals, which indicated that this fitness cost was recessive. Additionally, maize lines did not affect the magnitude of this fitness cost. The lower fitness of Cry1F‐resistant O. nubilalis may contribute to the maintenance of Cry1F susceptibility in field populations more than a decade after Cry1F maize was commercialized.     

Regulation of host diapause by an insect parasitoid

Abstract. 1. The interaction between larval development and parasitism by the braconid wasp Cotesia koebelei (Riley), was investigated in a population of the butterfly Euphydryas editha (Boisduval) (Nymphalidae). In this population, the butterfly host has an obligatory overwintering larval diapause.
2. It was found that E. editha larvae harbouring parasitoids were more likely to pass through an extra feeding instar before entering diapause than were non-parasitized conspecifics.
3. In addition, some individuals that were experimentally exposed to multiple parasitoid attacks bypassed diapause completely; these larvae passed through five or six feeding instars, reaching sizes typical of final instar post-diapause larvae.
4. The observed effect of superparasitism occurred regardless of whether the host larvae subsequently produced mature parasitoids, suggesting that parasitoid attack is sufficient to invoke the response.
5. It is proposed that the parasitoid C.koebelei regulates the number of pre-diapause feeding instars of its insect host E. editha, and that some component of the female venom, injected at oviposition, is responsible for this regulation.     

Effect of larval density on development and diapause of Sesamia nonagrioides (Lef.) (Lep., Noctuidae) under laboratory conditions

The effect of larval density on development and diapause of Sesamia nonagrioides (Lef.) was examined. Larvae were grown on an artificial diet at densities of five, 10, 20, 30 and 40 larvae per box, under two environmental (diapause-including and diapause-averting) conditions. High larval densities resulted in increased mortality under both experimental conditions tested. Although crowded larvae showed a higher pupation rate than those reared under low densities, small differences occurred between mean larval developmental time at different densities under developmental condition. In spite of this, crowding proved effective on the intensity of diapause of this species and the duration of diapause was shorter for crowded larvae. Pupal weights of both sexes were significantly reduced at high densities while there were significant differences between pupal weights formed by larvae reared under diapause conditions and those formed by larvae reared under nondiapause conditions at all density treatments. The pupal period was significantly longer under diapause conditions than that under development conditions but density was not found to play a role on the time of the emergence of the adults produced by larvae reared under developmental or diapause conditions.     

Carabid larvae as predators of weed seeds: granivory in larvae of Amara eurynota (Coleoptera: Carabidae)

Up to date we do not have much information about predation on seeds by larvae of ground beetles. One of the reasons why such knowledge is important is that granivorous larvae contribute to predation of weed seeds. In this study, the food requirements of larvae of autumn breeding carabid species Amara eurynota (Panzer) were investigated in the laboratory and a hypothesis, that they are granivorous was tested. Insect diet (Tenebrio molitor larvae), three seed diets (seeds of Artemisia vulgaris, Tripleurospermum inodorum or Urtica dioica or a mixed diet (T. molitor + A. uulgaris) were used as food. For larvae of A. eurynota, seeds are essential for successful completion of development, because all those fed pure insect diet died before pupation. However, differences in suitability were observed between pure seed diets. Larvae fed seeds of A. vulgaris had the lowest mortality and fastest development of the seed diets. Those fed seeds of T. inodorum had also low mortality, but the development was prolonged in the third instar. In contrast, development of larvae reared on seeds of U. dioica was slowest of the tested diets and could not be completed, as all individuals died before pupation. When insects were included to seed diet of A. vulgaris (mixed diet), the duration of development shortened, but mortality remained the same when compared to seed diet of A. vulgaris. According to the results it was concluded that larvae of A. eurynota are granivorous. A mixed diet and seed diets of A. uulgaris and T. inodorum were suitable and insect diet and seeds of U. dioica were unsuitable diets in this experiment.     

Composition of the Spruce Budworm (Choristoneura fumiferana) Midgut Microbiota as Affected by Rearing Conditions

The eastern spruce budworm (Choristoneura fumiferana) is one of the most destructive forest insect pests in Canada. Little is known about its intestinal microbiota, which could play a role in digestion, immune protection, communication and/or development. The present study was designed to provide a first characterization of the effects of rearing conditions on the taxonomic diversity and structure of the C. fumiferana midgut microbiota, using a culture-independent approach. Three diets and insect sources were examined: larvae from a laboratory colony reared on a synthetic diet and field-collected larvae reared on balsam fir or black spruce foliage. Bacterial DNA from the larval midguts was extracted to amplify and sequence the V6-V8 region of the 16S rRNA gene, using the Roche 454 GS-FLX technology. Our results showed a dominance of Proteobacteria, mainly Pseudomonas spp., in the spruce budworm midgut, irrespective of treatment group. Taxonomic diversity of the midgut microbiota was greater for larvae reared on synthetic diet than for those collected and reared on host plants, a difference that is likely accounted for by several factors. A greater proportion of bacteria from the phylum Bacteroidetes in insects fed artificial diet constituted the main difference between this group and those reared on foliage; within the phylum Proteobacteria, the presence of the genus Bradyrhizobium was also unique to insects reared on artificial diet. Strikingly, a Bray-Curtis analysis showed important differences in microbial diversity among the treatment groups, pointing to the importance of diet and environment in defining the spruce budworm midgut microbiota.     

Ontogenetic plasticity of food habits in the Mexican spiny-tailed iguana, Ctenosaura pectinata

Ontogenetic shifts from insect consumption by juveniles to plant consumption by adults are rare in the herbivorous lizard family Iguanidae. My investigations on diet and digestive tract anatomy of the iguanid lizard Ctenosaura pectinata show that this species has an ontogenetic diet shift. Insects were rare in adult diets but constituted 86.5% (by volume) of the food eaten by the smallest juveniles. All age classes ate some plant parts from a range of plant types, but flowers and leaves of legumes were a primarily food source. Non-adult lizards had the widest food niche breadths. Arthropods in the diet of juveniles and immatures covaried seasonally with the decline of arthropod abundance. Several hypotheses could explain this ontogenetic plasticity in diet. I rejected hypotheses that gut structure constrained juveniles to an arthropod diet and that insect consumption was purely an artifact of plant consumption because (1) size-adjusted gut morphology and capacity was similar among age classes, and (2) no food plants sampled had an excessive density of arthropods. I supported an alternative hypothesis that juveniles can eat plants but do not because insects provide a more nutritious diet. This conclusion was based on the observation that the juvenile hindgut is similar to that of herbivorous adults, and the propensity for juveniles to consume primarily, but not exclusively, insects when they were most abundant. The hindgut represents the site of fermentative plant fiber breakdown in many herbivorous lizards. Insect foods can compensate for size-related nutritional needs (energy and protein) and digestive limitations in juveniles. Opportunistic feeding to maintain a broad diet might help juvenile and immature lizards through high-predation-risk growth periods by reducing searching costs, increasing nutritional and energetic gains due to associative effects, and increasing new food exposure. Received: 20 January 1999 / Accepted: 25 January 2000     

Effects of submergence by winter floods on diapausing caterpillars of a wetland butterfly, Lycaena dispar batavus

1. The influence of seasonal flooding on wetland insects is understudied, and understanding of a potentially important influence on population dynamics is therefore incomplete. This is of particular consequence because many wetland insect species, including the large copper butterfly, Lycaena dispar batavus , are of conservation interest.
2. Previous studies on the submergence tolerance of diapausing L. d. batavus larvae have been inconclusive. This study investigated the effects of different periods of enforced submergence on overwintering survival, using both fresh and brackish water, and comparing effects on larvae in early and late diapause.
3. Larvae were submerged for up to 84 days, and survival was negatively correlated with submergence period, although periods of up to 28 days did not appear to reduce survival. Water type, i.e. fresh vs. brackish water, and larval stage, i.e. early vs. late diapause, had no significant influence on survival.
4. The relevance of these findings to the population biology and conservation of L. d. batavus is discussed. Further studies on the effects of submergence by seasonal flooding on wetland insect populations are encouraged.     

Timing of diapause initiation, metabolic changes and overwintering survival of the spruce budworm, Choristoneura fumiferana

1. Four groups of spruce budworm larvae, Choristoneura fumiferana , of the same physiological stage (at the beginning of diapause) were exposed to natural temperatures, starting in July, August, September and October. Post-diapause emergence and certain metabolites were monitored throughout the overwintering period.
2. Larval exposure to high temperatures for long periods before winter had a significant effect on temperature requirements for subsequent diapause development and apparently caused a certain amount of mortality, especially for those larvae that entered diapause earlier in the season.
3. Substantial loss of lipid and glycogen reserves was related to intensity of high temperature exposure during the pre-winter period, which in turn was associated with the timing of diapause initiation.
4. Patterns of glycerol accumulation during the winter were also influenced by temperature conditions in the early phase of diapause development.
5. Among several possible mortality factors, an extended cool period in spring was suggested as a potential cause of overwintering mortality, which was closely associated with a deficiency in energy reserves caused by high temperature exposure before winter.
6. These results stress the significant impact of pre-winter conditions on the overwintering process and highlight the importance of an ecophysiological approach to insect overwintering biology.     

A minimalist approach to the effects of density-dependent competition on insect life-history traits

Abstract  1. Due to its effects on the phenotypic and genotypic expression of life-history traits, density-dependent competition is an important factor regulating the growth of populations. Specifically for insects, density-dependent competition among juveniles is often associated with increased juvenile mortality, delayed maturity, and reduced adult size.
2. The aim of the work reported here was to test whether the established phenotypic effects of density-dependent competition on life-history traits could be reproduced in an experimental design requiring a minimal number of individuals. Larvae of the mosquito Aedes aegypti were reared at densities of one, two, or three individuals per standard Drosophila vial and in six different conditions of larval food availability. This design required relatively few individuals per independent replicate and included a control treatment where individuals reared at a density of one larva per vial experienced no density-dependent interactions with other larvae.
3. Increased larval densities or reduced food availability led to increased larval mortality, delayed pupation, and the emergence of smaller adults that starved to death in a shorter time (indicating emergence with fewer nutritional reserves).
4. Female mosquitoes were relatively larger than males (as measured by wing length) but males tended to survive for longer. These differences increased as larval food availability increased, indicating the relative importance of these two traits for the fitness of each sex. The role of nutritional reserves for the reproductive success of males was highlighted in particular.
5. This minimalist approach may provide a useful model for investigating the effects of density-dependent competition on insect life-history traits.